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Detection of functional groups and antibodies on microfabricated surfaces by confocal microscopy

Journal Article · · Biotechnology and Bioengineering
;  [1];  [2]
  1. Univ. of California, Berkeley, CA (United States)
  2. Lawrence Berkeley National Lab., CA (United States)

Fluorescence confocal microscopy was used to characterize micron-sized microfabricated silicon particles and planar oxides surfaces after silanization and immobilization of IgG antibody. Surfaces treated with amino- and mercaptosilanes were tested by the presence of amine and sulfhydryl groups by labeling with specific fluorescein probes. In addition, human antibody (IgG) was immobilized to the thiol-coated microparticles using the heterobifunctional crosslinker succinimidyl 4-(N-maleimidolmthyl)-cyclohexane-1-carboxylate. Estimates of the surface density of IgG were consistent with 8.3% of a monolayer of covalently-bound antibody. Confocal images confirmed uniform layers of both silanes and antibodies on the microparticles. The sensitivity limit for the confocal measurements was determined to be as low as 1.5 x 10{sup {minus}5} fluors per nm{sup 2}.

Sponsoring Organization:
USDOE, Washington, DC (United States)
DOE Contract Number:
AC03-76SF00098
OSTI ID:
290241
Journal Information:
Biotechnology and Bioengineering, Journal Name: Biotechnology and Bioengineering Journal Issue: 2 Vol. 60; ISSN BIBIAU; ISSN 0006-3592
Country of Publication:
United States
Language:
English

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